The role of myosin light chain kinase in intestinal epithelial barrier dysfunction due to hypoxia
Author(s): WANG Pei, CHEN Chuan-li, Li Mu, WANG Feng-jun
Pages: 57-60
Year: 2009 Issue: 1
Journal: Chinese Journal of Burns
Keyword: Anoxia; Myosin light chain; Myosin light chain kinase; Intestinal epithelial cell; Barrier function;
Abstract: Objective To study the role of myosin light chain kinase (MLCK) in intestinal epithe-lial barrier dysfunction after hypoxia. Methods The Caco-2 monolayers developed with Transwell inserts were exposed to hypoxia for 0 h(NC group), 2, 6, 8, 12 and 24 h(H group), and 6 h hypoxic specimens were treated with 100 mul/L ML-9 (T group). The transepithelial electrical resistance (TER) of monolayers was measured with an ohmmeter. The tight junction protein ZO-1 of monolayers was analyzed by immunofluo-rescence assay. The protein expressions of phosphorylated myosin light chain (p-MLC) and MLCK were de-tected by Western blotting. Results The TER of monolayers in H group at 6, 8, 12 and 24 h was 422± 17, 427±27,403±40 and 426±22Ω respectively, which was significantly lower than that of NC group (451±27Ω, P < 0.05). The TER of monulayers in T group was 558±110Ω, which was significantly high-er than that in H group at each time point (P <0.01). The ZO-1 of monolayers in H group at 6 h was ir-regular in arrangement, with interruptions and rugae, and sawtooth. These abnormalities were amelioated in T group(regular in arrangement, with little or without ruga and sawtooth). The protein expressions of p-MLC and MLCK in H group at each time point were higher than those in NC group. Conclusions Intestinal ep-ithelial barrier dysfunction after hypoxia can be mediated by MLCK.
Pages: 57-60
Year: 2009 Issue: 1
Journal: Chinese Journal of Burns
Keyword: Anoxia; Myosin light chain; Myosin light chain kinase; Intestinal epithelial cell; Barrier function;
Abstract: Objective To study the role of myosin light chain kinase (MLCK) in intestinal epithe-lial barrier dysfunction after hypoxia. Methods The Caco-2 monolayers developed with Transwell inserts were exposed to hypoxia for 0 h(NC group), 2, 6, 8, 12 and 24 h(H group), and 6 h hypoxic specimens were treated with 100 mul/L ML-9 (T group). The transepithelial electrical resistance (TER) of monolayers was measured with an ohmmeter. The tight junction protein ZO-1 of monolayers was analyzed by immunofluo-rescence assay. The protein expressions of phosphorylated myosin light chain (p-MLC) and MLCK were de-tected by Western blotting. Results The TER of monolayers in H group at 6, 8, 12 and 24 h was 422± 17, 427±27,403±40 and 426±22Ω respectively, which was significantly lower than that of NC group (451±27Ω, P < 0.05). The TER of monulayers in T group was 558±110Ω, which was significantly high-er than that in H group at each time point (P <0.01). The ZO-1 of monolayers in H group at 6 h was ir-regular in arrangement, with interruptions and rugae, and sawtooth. These abnormalities were amelioated in T group(regular in arrangement, with little or without ruga and sawtooth). The protein expressions of p-MLC and MLCK in H group at each time point were higher than those in NC group. Conclusions Intestinal ep-ithelial barrier dysfunction after hypoxia can be mediated by MLCK.
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